CN102613384A - Method for preparing spiral seaweed polypeptide powder by using living spiral seaweeds - Google Patents
Method for preparing spiral seaweed polypeptide powder by using living spiral seaweeds Download PDFInfo
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Abstract
The invention discloses a method for preparing spiral seaweed polypeptide powder by using living spiral seaweeds. The conventional method for preparing the spiral seaweed polypeptide is usually to prepare a suspension by using dry spiral seaweed powder and water and then hydrolyze the spiral seaweed protein to prepare the spiral seaweed polypeptide by using biological enzyme engineering technology, but the final product has insufficient activity and low extraction rate as plenty of active components of the dry spiral seaweed powder are lost, and fishy smell of the spiral seaweed cannot be removed. The method for preparing spiral seaweed polypeptide powder by using the living spiral seaweeds, disclosed by the invention, comprises the step of preparing the spiral seaweed polypeptide powder through three-step enzyme hydrolysis method of alkali protease, flavourzyme and monascus by adopting the living spiral seaweeds as the raw material. The preparing spiral seaweed polypeptide powder by using the living spiral seaweeds, disclosed by the invention, has the advantages of high extraction rate, being capable of keeping the active components of the spiral seaweed polypeptide products efficiently, preparing the piral seaweed polypeptide without fishy smell and having better taste.
Description
Technical field
The present invention relates to a kind of method for preparing the spirulina polypeptide powder with the live body spirulina.
Background technology
The high-quality high protein content is 3 times of fish up to 60～70% in the spiral frond, is 4 times of meat, be more than 5 times of egg, and chlorophyll content is more than 10 times of vegetables.In addition, the content of vitamin B3 approximately is 6.4 times of soybean; Beta carotene approximately is 4 times of content in the common carrot; Approximately contain 11 kinds of aliphatic acid in the spirulina, linoleic acid plus linolenic acid wherein all is an essential fatty acid; Spirulina contains essential constant of humans and animals and trace element, not only contains a large amount of iron, and can form organic iron with protein and algocyan chelating, thereby helps being absorbed and used; Spirulina has the ability of enrichment selenium, zinc, iodine simultaneously.Spirulina is a kind of desirable nutrient and healthcare products, and market prospects are fine.
There is following problem in the application of spirulina at present: one of which, spirulina have denseer fishy smell, and taste is relatively poor.They are two years old; Absorb for strengthening; Spirulina will be prepared into spirulina polypeptide usually, prepares spirulina polypeptide at present and typically uses spirulina powder and add water and be mixed with suspension, prepares spirulina polypeptide with bio-enzyme engineering technology hydrolysis spirulina protein again; But the spirulina powder active ingredient loses in a large number, makes final products active not enough.Its three, the method enzymolysis degree that existing general bio-enzyme engineering technology hydrolysis spirulina protein prepares spirulina polypeptide is lower, effectively enzymolysis time is shorter, recovery rate is lower, wastage of material is serious.
In the prior art; The preparation method of the disclosed spirulina polypeptide of CN101095458A; It has comprised that dry powder obtain solution, neutral proteinase take off steps such as raw meat, breaking-wall cell, enzymatic hydrolysis (cellulase and pectase, papain), centrifugal filtration, concentrate drying, adopts enzymolysis (neutral proteinase) and physical treatment (beta-schardinger dextrin-parcel) way of combining to take off raw meat.In addition; CN1204651A discloses the preparation method of spirulina polypeptide and has contained the medicament and the application of these polypeptide; Said method is used spirulina powder; In acid, alkalescence or neutral ring mirror,, and combine multiple hydrolysising protease to obtain spirulina polypeptide by magnesium, calcium, phosphorus, selenium plasma hydrolysis catalysis.All adopting spirulina powder in the said method is raw material, selects clasmatosis to extract protein.
Summary of the invention
The purpose of this invention is to provide and a kind ofly prepare the method for spirulina polypeptide powder with the live body spirulina, having solved existing spirulina powder, to extract spirulina polypeptide method recovery rate low, and the finished product content of peptides is few, the shortcoming of taste bad will.The method recovery rate is high, can effectively keep spirulina polypeptide product activity composition, and the spirulina polypeptide of the method preparation does not simultaneously have fishy smell, and taste is better.
This method is with respect to the preparation method's of spirulina polypeptide in the prior art advantage: it is the feedstock production spirulina polypeptide that the live body spirulina is adopted in (1); Adopting spirulina powder compared with prior art is raw material; Can increase the activity of final products; And on preparation technology, can reduce the step of pre-treatment, enhance productivity; (2) taking off in the raw meat processing; Unexpected discovery; Adopt flavor protease to can solve the problem that spirulina polypeptide fishy smell reaches taste bad will greatly,, be more conducive to produce extension than the technology with the beta-schardinger dextrin-parcel is more easy again behind the available technology adopting enzymolysis; (3) select monascus to be used for the enzymolysis spirulina, the significant prolongation enzymolysis time makes algae mud enzymolysis abundant, improves recovery rate.(4) select optimum enzymolysis scheme, promptly make up degree of hydrolysis, and confirmed the parameter of optimum enzymolysis test through test with the raising spirulina polypeptide through alkali protease, flavor protease, monascus.
Technical scheme of the present invention is following:
(1) the live body spirulina filters cleaning, draining is handled;
The live body spirulina is filtered cleaning, behind the adjusting pH algae mud is carried out draining and handle, make the water content of algae mud be less than 90%.
(2) first step enzymolysis: alkali protease;
The algae mud of handling well is packed in the retort, add alkali protease, accomplish first step enzyme digestion reaction.
(3) second step enzymolysis: flavor protease;
After first step enzymolysis is accomplished, add flavor protease, accomplish the second step enzyme digestion reaction.
(4) the 3rd step enzymolysis: monascus;
After the second step enzymolysis is accomplished, add monascus, accomplish the 3rd step enzyme digestion reaction.
(5) spray-drying.
Use the spray drying tower spray-drying, gained is live body spirulina polypeptide powder.
Estimating the normal index that adopts of spirulina polypeptide preparation method in the prior art is the degree of hydrolysis of spirulina protein; The present invention is high except recovery rate with respect to the preparation method's of spirulina polypeptide in the prior art advantage; Outside the degree of hydrolysis height; Because adopting the live body spirulina is the feedstock production spirulina polypeptide, adopting spirulina powder compared with prior art is raw material, can increase the activity of final products.Therefore, the present invention also introduces the mensuration of the activity of spirulina polypeptide.
(1) experimental technique
1, the mensuration of spirulina protein degree of hydrolysis:
Determination of Hydrolysis Degree of Protein adopts pH-stat method (Adler-Nissen, 1986),
Spirulina protein degree of hydrolysis (DH) accounts for generic key number (h with the peptide number of keys (h) of hydrolytic cleavage
Tot) percentage confirm, that is:
DH＝h/h
tot×100％
2, the mensuration of the activity of spirulina polypeptide
Phycocyanin is a kind of natural algocyan albumen of biologically active function in the spirulina, is active material important in the spirulina.Therefore, the content of measuring phycocyanin in the spirulina polypeptide can reflect the activity of spirulina polypeptide preferably.
The mensuration of phycocyanin concentration is with reference to [Qu Wenjuan etc., the impulse ultrasound auxiliary extraction technology of blunt top spirulina azurin; Food Science, 2007 5 phases, 135-138] in method; The characteristic absorption of phycocyanin is at 620nm, and the characteristic absorption of allophycocyanin is at 650nm.Phycocyanin concentration (C) (I) calculating by formula in the extract:
C＝(A
620nm-0.474A
650nm)/5.34，(mg/ml)
3, different enzymolysis prescriptions are to the influence of spirulina protein degree of hydrolysis
Zymyhydrolyzed protein matter is the main path that obtains polypeptide at present, and this method advantage is saving cost, Product Safety height, has no side effect.Hydrolysis is the key of producing spirulina polypeptide with the selection of enzyme.
This experiment has selected for use common business-like protease and monascus to handle spirulina, all is employed in 35 ℃, hydrolysis 60,90,120,180 minutes.Used protease of the present invention and monascus are by buy on the market, and protease comprises alkali protease (enzyme activity is 400,000 U/g), flavor protease (enzyme activity is 1.5 ten thousand U/g), papain.The result sees table 1.
The different enzymolysis prescriptions of table 1 are to the influence of spirulina protein degree of hydrolysis (DH (%))
Visible by table 1, wherein best to the hydrolysis effect of spirulina protein with alkali protease.Therefore, the present invention at first selects the hydrolysis by novo spirulina for use; And further select flavor protease to remove the peculiar fishy smell of spirulina, significantly improve the taste of spirulina polypeptide; In addition, consider monascus ability significant prolongation enzymolysis time, make algae mud enzymolysis abundant, improve recovery rate.Based on above consideration, the present invention finally confirms to come enzymolysis live body spirulina with three step enzymatic isolation methods of alkali protease, flavor protease, monascus combination.
4, three step enzymatic isolation method process parameter optimizings
(1) single factor experiment influences degree of hydrolysis
The pH value is one of essential condition of enzymatic reaction, and suitable pH value is the important parameter of enzymatic reaction.It is to be hydrolyzed under 6,6.5,7,7.5,8,8.5 the condition that experiment is located at the pH value, and hydrolysis time is 3 hours.The result sees table 2:
The different pH values of table 2 are to the influence of spirulina protein degree of hydrolysis (DH (%))
Visible by table 2, degree of hydrolysis increases along with the pH value and raises basically.After taking all factors into consideration, confirm that selecting the pH value is the 8.5 pH values as enzyme digestion reaction.
In the enzyme optimum temperature range, choose 6 levels such as 35 ℃ of hydrolysis temperatures, 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃, investigate protein degree after the enzyme digestion reaction.The result sees table 3:
Table 3 different temperatures is to the influence of spirulina protein degree of hydrolysis (DH (%))
Visible by table 3, enzyme and most of chemical reaction have points of resemblance basically, and under lower temperature, degree of hydrolysis constantly rose when temperature raise, but when temperature continuation rising, zymoprotein divides sex change and inactivation, cause that enzyme reaction speed descends.Therefore, finally the temperature of definite enzyme reaction is 45-50 ℃.
(2) multifactorial experiment influences degree of hydrolysis
Confirmed that through above-mentioned experiment the pH value of the best enzyme digestion reaction of corresponding alkali protease, flavor protease and monascus is 8.5 in the three step enzymatic isolation methods, temperature is 45-50 ℃.
Next, under the condition that limits pH value and temperature, confirm enzyme concentration and enzyme digestion reaction time.
Refer to alkali protease, flavor protease and monascus with J, F, D respectively.The result sees table 4
Table 4 multifactorial experiment influences degree of hydrolysis
Visible by table 4, best enzyme concentration and enzymolysis time is respectively in the three step enzymatic isolation methods: in the weight of algae mud, and alkali protease (0.05%), flavor protease (0.03%) and monascus (0.15%); Alkali protease (70min), flavor protease (60min) and monascus (180min).
Through above test, confirm that finally three of the best of the present invention goes on foot the experimental technique of enzymatic isolation methods, be the most preferred technical scheme of the present invention to be:
(1) the live body spirulina is filtered cleaning, pH reaches at 8.5 o'clock algae mud is carried out the draining processing, makes the water content of algae mud be less than 90%.Obtain algae mud after the processing.
(2) first step enzymolysis:
The algae mud of handling well is packed in the retort, add alkali protease, enzyme activity is 400,000 U/g, very calculates according to quality, and alkali protease is 0.05% of an algae mud, and hydrolysis temperature is 45-50 ℃, and be 70 minutes action time, and first step enzymolysis is accomplished.
(3) second step enzymolysis:
After first step enzymolysis is accomplished, add flavor protease, enzyme activity is 1.5 ten thousand U/g, very calculates according to quality, and flavor protease is 0.03% of an algae mud, and hydrolysis temperature is 45-50 ℃, and be 60 minutes action time, and the second step enzymolysis is accomplished.
(4) the 3rd step enzymolysis:
After the second step enzymolysis is accomplished, add monascus, very calculate according to quality, monascus is 0.15% of an algae mud, and operative temperature is 45-50 ℃, and be 180 minutes action time.
(5) spray-drying: use the spray drying tower spray-drying, gained is live body spirulina polypeptide powder.
(2) specific embodiment
The live body spirulina is filtered cleaning, and pH reaches at 8.5 o'clock algae mud is carried out the draining processing, and the water content that makes algae mud is less than 90%.The algae mud of handling well is packed in the retort, and 1000 kilograms every jar, adding enzyme activity is 0.5 kilogram of 400,000 U/g alkali protease, heats, stirs, and hydrolysis temperature is 45 ℃, and be 70 minutes action time.Add enzyme activity and be 0.3 kilogram of the flavor protease of 1.5 ten thousand U/g, heat, stir, hydrolysis temperature is 45 ℃, and be 60 minutes action time.Add 1.5 kilograms of monascuses, heat, stir, 180 minutes use spray drying tower spray-dryings of reaction under 45 ℃ of conditions, gained is live body spirulina polypeptide powder.
The activity of spirulina polypeptide and Determination of Hydrolysis Degree of Protein.
With reference to " in (one) experimental technique ", the assay method of the activity of spirulina polypeptide, measure the activity of the spirulina polypeptide of prepared spirulina polypeptide powder among the embodiment.
As a result, phycocyanin content is 16%, and the activity of the spirulina polypeptide that visible the present invention makes is significantly higher than the activity (about 5-9%) of the spirulina polypeptide that conventional method makes.
According to above-mentioned protein degree assay method, record the protein degree 38 (%) of spirulina polypeptide of the present invention, the degree of hydrolysis of the spirulina polypeptide that visible the present invention makes is significantly higher than the protein degree (about 10-20%) that conventional method obtains.
Claims (2)
1. one kind prepares the method for spirulina-active polypeptide powder with the live body spirulina, it is characterized in that choosing the live body spirulina, carries out the following operating procedure:
(1) the live body spirulina filters cleaning, draining is handled:
The live body spirulina is filtered cleaning, behind the adjusting pH algae mud is carried out draining and handle, make the water content of algae mud be less than 90%,
(2) first step enzymolysis:
The algae mud of handling well is packed in the retort, adds alkali protease, accomplish first step enzyme digestion reaction,
(3) second step enzymolysis:
After first step enzymolysis is accomplished, add flavor protease, accomplish the second step enzyme digestion reaction,
(4) the 3rd step enzymolysis:
After the second step enzymolysis is accomplished, add monascus, accomplish the 3rd step enzyme digestion reaction,
(5) spray-drying:
Use the spray drying tower spray-drying.
2. the preparation method of the described spirulina-active polypeptide powder of claim 1 is characterized in that said operations step is:
(1) the live body spirulina filter is cleaned, PH reaches at 8.5 o'clock to carry out draining with algae mud and handles, and the water content that makes algae mud is less than 90%,
(2) the algae mud of handling well is packed in the retort, add alkali protease, enzyme activity is 400,000 U/g, very calculates according to quality, and alkali protease is 0.05% of an algae mud, heats, stirs, and hydrolysis temperature is 45-50 ℃, and be 70 minutes action time,
(3) add flavor protease, enzyme activity is 1.5 ten thousand U/g, very calculates according to quality, and flavor protease is 0.03% of an algae mud, heats, stirs, and hydrolysis temperature is 45-50 ℃, and be 60 minutes action time,
(4) add monascus, very calculate according to quality, monascus is 0.15% of an algae mud, and temperature is 45-50 ℃, and be 180 minutes action time,
(5) use the spray drying tower spray-drying.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104432079A (en) * | 2014-12-29 | 2015-03-25 | 陕西天宝大豆食品技术研究所 | Whole-seaweed-peptide nourishment and preparation method thereof |
| CN105248837A (en) * | 2015-11-16 | 2016-01-20 | 岳军堂 | Chlorella active polypeptide powder preparing method |
| CN110346387A (en) * | 2019-06-29 | 2019-10-18 | 浙江大学 | A method of trichome of spirulina draining performance is identified using transmission electron microscope |
| CN110463900A (en) * | 2019-07-08 | 2019-11-19 | 华南农业大学 | A method of removing fish fermented product fishy smell |
| CN110904178A (en) * | 2019-12-03 | 2020-03-24 | 襄阳蒙肽生物有限公司 | Preparation method of protein polypeptide |
| CN110999933A (en) * | 2019-12-03 | 2020-04-14 | 襄阳蒙肽生物有限公司 | Polypeptide biscuit and preparation method thereof |
| CN111154824A (en) * | 2020-01-15 | 2020-05-15 | 润科生物工程（福建）有限公司 | Industrial production method for obtaining spirulina antioxidant oligopeptide through high-concentration two-step enzymolysis |
| CN112998279A (en) * | 2021-03-15 | 2021-06-22 | 东北农业大学 | Method for preparing functional polypeptide powder by enzyme method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2857978A1 (en) * | 2003-07-25 | 2005-01-28 | Simer Laboratoires Science Et | Preparing a peptide extract of Spirulina, useful in nutraceutical and cosmetic compositions for e.g. controlling aging of the skin, comprises extraction of lipids then enzymatic hydrolysis |
| CN101095458A (en) * | 2007-07-19 | 2008-01-02 | 武汉天天好生物制品有限公司 | Method for preparation of spirulina polypeptide |
| CN101978888A (en) * | 2010-10-25 | 2011-02-23 | 广东润科生物工程有限公司 | Method for preparing microalgae polypeptide powder and microalgae polypeptide powder-containing flavoring |
| US20110218326A1 (en) * | 2010-03-03 | 2011-09-08 | Zen-U Biotechnology Co., Ltd. | Plant-protein product and method for preparing the same |
-
2012
- 2012-04-18 CN CN 201210113966 patent/CN102613384B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2857978A1 (en) * | 2003-07-25 | 2005-01-28 | Simer Laboratoires Science Et | Preparing a peptide extract of Spirulina, useful in nutraceutical and cosmetic compositions for e.g. controlling aging of the skin, comprises extraction of lipids then enzymatic hydrolysis |
| CN101095458A (en) * | 2007-07-19 | 2008-01-02 | 武汉天天好生物制品有限公司 | Method for preparation of spirulina polypeptide |
| US20110218326A1 (en) * | 2010-03-03 | 2011-09-08 | Zen-U Biotechnology Co., Ltd. | Plant-protein product and method for preparing the same |
| CN101978888A (en) * | 2010-10-25 | 2011-02-23 | 广东润科生物工程有限公司 | Method for preparing microalgae polypeptide powder and microalgae polypeptide powder-containing flavoring |
Non-Patent Citations (2)
| Title |
|---|
| 刘士伟: "螺旋藻降解肽的分离制备及其抑菌性的研究", 《中国优秀硕士学位论文全文数据库工程科技I辑》 * |
| 杨磊，等: "酶解螺旋藻泥制备活性肽的工艺条件优化研究", 《食品科技》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104432079A (en) * | 2014-12-29 | 2015-03-25 | 陕西天宝大豆食品技术研究所 | Whole-seaweed-peptide nourishment and preparation method thereof |
| CN105248837A (en) * | 2015-11-16 | 2016-01-20 | 岳军堂 | Chlorella active polypeptide powder preparing method |
| CN110346387A (en) * | 2019-06-29 | 2019-10-18 | 浙江大学 | A method of trichome of spirulina draining performance is identified using transmission electron microscope |
| CN110463900A (en) * | 2019-07-08 | 2019-11-19 | 华南农业大学 | A method of removing fish fermented product fishy smell |
| CN110904178A (en) * | 2019-12-03 | 2020-03-24 | 襄阳蒙肽生物有限公司 | Preparation method of protein polypeptide |
| CN110999933A (en) * | 2019-12-03 | 2020-04-14 | 襄阳蒙肽生物有限公司 | Polypeptide biscuit and preparation method thereof |
| CN111154824A (en) * | 2020-01-15 | 2020-05-15 | 润科生物工程（福建）有限公司 | Industrial production method for obtaining spirulina antioxidant oligopeptide through high-concentration two-step enzymolysis |
| CN112998279A (en) * | 2021-03-15 | 2021-06-22 | 东北农业大学 | Method for preparing functional polypeptide powder by enzyme method |
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